Virus Handling and Storage

Recombinant viruses are replication deficient and can infect a broad range of mammalian cell types. BrainVTA recommends the following steps to ensure proper handling and storage for virus. If there is any problem, please contact BrainVTA within 10 days of receiving your virus. If your virus did not arrive on dry ice, please contact BrainVTA immediately.

Safety

• Adeno-Associated Virus (AAV)
  Recombinant AAV constructs do not encode for either a potentially tumorigenic gene product or a toxin molecule. According to guidelines from the National Institutes of Health (NIH), recombinant AAV vectors can be handled in a Biosafety Level 1 (BSL-1). If dealing with biohazardous material then please handle under Biosafety Level 2 (BSL-2) containment.
• Lentivirus (LV)
  Lentiviral vector constructs are derived from HIV and are therefore highly efficient vehicles for in vivo gene delivery. They can integrate transgenes into dividing and non-dividing cells, so work with lentiviral vectors must be carefully. NIH guidelines require the maintenance of a BSL-2 or BSL-2 enhanced control facility for work involving lentivirus.

This is the conventional guide for using, please also refer to your institution's Occupational Health and Safety office for guidance on safe handling of viral particles.

Storage

Lentivirus and AAV should be stored immediately at -80°C after receipt. For short-term storage, all vectors can be stored at room temperature or 4°C. For long-term storage, keep all vectors at -80°C. Do NOT store at -20°C. Research suggests that long-term storage at -20°C may result in decreased transduction efficiency of the virus.

We recommend aliquoting your virus into desired volumes, snap-freezing in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C for use in future experiments. Please avoid unnecessary freeze-thawing of the vectors, as excessive freeze-thawing cycles may result in a significant decrease in titer and loss of biological activity.

Thawing and Handling Your Virus

• Thawing prior to use: On ice or at room temperature, and use immediately.
• During experiment: Keep on ice at all times.

Cell Transduction

AAV and Lentivirus particles from both small scale and large scale packaging can be applied directly for cell transduction. Lentiviruses can infect most cells in vitro, while AAV is not as efficient as Lentiviruses in vitro. Please review the literature and perform pre-experiments before the formal experiment to find the best MOI.

Animal Injections

All virus provided is dissolved in F68/PBS, and is suitable for in vivo use. Please review the literature and perform pre-experiments before the formal experiment. AAV viral tropism is determined by many factors and will vary between cell lines or host tissues. Therefore, when choosing AAV, pay attention to choosing the right serotype and promoter. If you have any questions in the selection process, please feel free to contact us at sales@brainvta.com!